Basis of Mutual Domain Inhibition in a Bacterial Response Regulator

Corrêa, Fernando Morgan de Aguiar; Gardner, Kevin H.

doi:10.1016/j.chembiol.2016.07.010

Cited by 17 publications

(20 citation statements)

References 50 publications

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“…RMSF reports on the variance in the position of each residue, and was particularly elevated in the α4 loop after restructuring (Supporting Information Fig. S1B and C); we also observed some modest fluctuation in the region surrounding the phosphorylation site that is consistent with increased dynamics as a function of phosphorylation (Campagne et al ., ; Herrou et al ., ; Corrêa and Gardner, ).…”

Section: Resultsmentioning

confidence: 99%

“…). NMR studies of E. litoralis PhyR (56% identical to C. crescentus PhyR; BLAST e < 3e −96 ) show widespread changes in chemical shifts as a function of modification with the phosphomimetic compound, beryllium fluoride (

{BeF}_{3}^{-}

) (Corrêa and Gardner, ). Thus, PhyR phosphorylation clearly alters PhyR structure.…”

Section: Resultsmentioning

confidence: 99%

“…T220 is highly conserved and stabilizes the phosphoryl group in REC domains that utilize either Y‐T coupling or FATGY signaling mechanisms (Cho et al ., ; Sheftic et al ., ; Campagne et al ., ). This residue is required for conformational changes induced by

{BeF}_{3}^{-}

modification in E. litoralis PhyR (Corrêa and Gardner, ). The structural role of R225 is unclear, though we note that R225 stacks with F222 and may provide stability to the α11 helix.…”

Section: Resultsmentioning

confidence: 99%

“…We provide evidence that phosphorylation of the PhyR REC domain is not sufficient to induce formation of a large fraction of open‐state PhyR in vitro , though disruption of a hydrogen bonding network in the α11‐β5 loop and solvation of residues at the REC‐SL interface (upon phosphorylation) are associated with a lowered energetic barrier to opening (Figs ). This result is consistent with a growing body of data that implicate α11‐β5 conformational change in PhyR activation (Campagne et al ., ; Herrou et al ., ; Corrêa and Gardner, ,). Our data support a GSR activation model in which NepR interaction with PhyR, likely via an intrinsically disordered region of NepR polypeptide, supplies free energy to open and activate phospho‐PhyR.…”

Section: Discussionmentioning

confidence: 99%

“…Recent nuclear magnetic resonance (NMR) spectroscopy studies of full‐length Erythrobacter litoralis PhyR and its isolated REC domain demonstrate that (i) aspartyl modification of the PhyR REC domain by the phosphomimetic compound

{BeF}_{3}^{-}

shifts REC toward an active conformation in full‐length protein and (ii) the isolated PhyR REC domain adopts an active conformation even in the absence of

{BeF}_{3}^{-}

modification (Corrêa and Gardner, ). Thus, the PhyR REC domain is held in an inactive conformation by its interaction with the SL domain on the same polypeptide.…”

Section: Introductionmentioning

confidence: 99%

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Allosteric control of a bacterial stress response system by an anti‐σ factor

Luebke

Eaton

Sachleben

et al. 2017

Molecular Microbiology

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Bacterial signal transduction systems commonly use receiver (REC) domains, which regulate adaptive responses to the environment as a function of their phosphorylation state. REC domains control cell physiology through diverse mechanisms, many of which remain understudied. We have defined structural features that underlie activation of the multi-domain REC protein, PhyR, which functions as an anti-anti-σ factor and regulates transcription of genes required for stress adaptation and host-microbe interactions in Alphaproteobacteria. Though REC phosphorylation is necessary for PhyR function in vivo, we did not detect expected changes in inter-domain interactions upon phosphorylation by solution X-ray scattering. We sought to understand this result by defining additional molecular requirements for PhyR activation. We uncovered specific interactions between unphosphorylated PhyR and an intrinsically disordered region (IDR) of the anti-σ factor, NepR, by solution NMR spectroscopy. Our data support a model whereby nascent NepR(IDR)-PhyR interactions and REC phosphorylation coordinately impart the free energy to shift PhyR to an open, active conformation that binds and inhibits NepR. This mechanism ensures PhyR is activated only when NepR and an activating phosphoryl signal are present. Our study provides new structural understanding of the molecular regulatory logic underlying a conserved environmental response system.

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Section: Resultsmentioning

confidence: 99%

{BeF}_{3}^{-}

) (Corrêa and Gardner, ). Thus, PhyR phosphorylation clearly alters PhyR structure.…”

Section: Resultsmentioning

confidence: 99%

{BeF}_{3}^{-}

modification in E. litoralis PhyR (Corrêa and Gardner, ). The structural role of R225 is unclear, though we note that R225 stacks with F222 and may provide stability to the α11 helix.…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

{BeF}_{3}^{-}

shifts REC toward an active conformation in full‐length protein and (ii) the isolated PhyR REC domain adopts an active conformation even in the absence of

{BeF}_{3}^{-}

modification (Corrêa and Gardner, ). Thus, the PhyR REC domain is held in an inactive conformation by its interaction with the SL domain on the same polypeptide.…”

Section: Introductionmentioning

confidence: 99%

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